Plant Growth Regulation

, Volume 51, Issue 1, pp 11–19 | Cite as

The dynamics of nutrient utilization and growth of apple root stock ‘M9 EMLA’ in temporary versus continuous immersion bioreactors

  • D. Chakrabarty
  • Y. H. Dewir
  • E. J. Hahn
  • S. K. Datta
  • K. Y. Paek
Original Paper


The present study investigated the dynamics of nutrient utilization and various growth and physiological parameters during in vitro proliferation of apple root stock ‘M9 EMLA’ in two different bioreactor systems, i.e. temporary and continuous immersions. Individual shoots obtained from temporary immersion system had higher dry mass and were of better quality than those obtained from continuous immersion. In continuous immersion bioreactor, apple shoots appeared to utilize more nutrients from liquid culture medium than that from temporary immersion. The shoot growth was limited by the availability of phosphate and nitrogen in continuous immersion system. The shoots produced in temporary immersion bioreactor showed higher photosynthetic rate, maximum quantum yield of photosystem-II and slow but steady rate of nutrient absorption, indicating the occurrence of higher photomixotrophic metabolism. The study also showed that high level of antioxidant scavenging enzymes in shoots grown in continuous immersion system induced physiological changes to foster adaptation to stresses.


Antioxidant enzymes Apple Bioreactor Continuous immersion system Fv/Fm Nutrient utilization Temporary immersion system 



Ascorbate peroxidase


Balloon-type bubble bioreactor






Indole-3-butyric acid


Guaiacol peroxidase


Gibberellic acid


Glutathione reductase


Murashige and Skoog’s (1962) medium


Photosynthetic photon flux density


Reactive oxygen species


Superoxide dismutase



This work was financially supported by the Ministry of education and Human Resource Development (MOE), the Ministry of Commerce, Industry and Energy (MOCIE) and the Ministry of Labor (MOLAB) through the fostering project of the Lab of Excellency. DC acknowledges the KOSEF for providing financial assistance in the form of “Long-term Foreign Scientist Program”. SKD also acknowledges the financial assistance from INSA-KOSEF Exchange Program.


  1. Aebi H (1974) Catalases. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 673–684Google Scholar
  2. Ahmed S, Nawata E, Hosokawa M, Domae Y, Sakuratani T (2002) Alterations in photosynthesis and some antioxidant enzymatic activities of mungbean subjected to waterlogging. Plant Sci 163:117–123CrossRefGoogle Scholar
  3. Aitken-Christie J (1991) Automation. In: Debergh PC, Zimmerman RJ (eds) Micropropagation: technology and application. Kluwer Academic Publishers, Dordrecht, pp 363–388Google Scholar
  4. Alvard D, Cote F, Teisson C (1993) Comparison of methods of liquid medium culture for banana micropropagation. Plant Cell Tissue Organ Cult 32:55–60CrossRefGoogle Scholar
  5. Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in condition. Anal Biochem 161:559–566PubMedCrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 PubMedCrossRefGoogle Scholar
  7. Chakrabarty D, Hahn EJ, Yoon YS, Paek KY (2003) Micropropagation of apple root stock ‘M9 EMLA’ using bioreactor. J Hortic Sci Biotechnol 78:605–609Google Scholar
  8. Chakrabarty D, Park SY, Ali MB, Shin KS, Paek KY (2006) Hyperhydricity in apple: physiological and ultrastructural aspects. Tree Physiol 26:377–388PubMedGoogle Scholar
  9. Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. Plant Cell Physiol 30:987–998Google Scholar
  10. Debergh PC, Read PE (1990) Micropropagation. In: Debergh PC, Zimmerman RH (eds) Micropropagation: technology and application. Kluwer Academic Publishers, Dordrecht, pp 1–13Google Scholar
  11. Dewir YH, Chakrabarty D, Hahn EJ, Paek KY (2005) Reversion of inflorescence in Euphorbia milii and its application to large scale micropopagation in an air-lift bioreactor. J Hortic Sci Biotechnol 80:581–587Google Scholar
  12. Etienne H, Berthouly M (2002) Temporary immersion systems in plant micropropagation. Plant Cell Tissue Organ Cult 69:215–231CrossRefGoogle Scholar
  13. Etienne H, Lartaud M, Michaux-Ferrière N, Carron MP, Berthouly M, Teisson C (1997) Improvement of somatic embryogenesis in Hevea brasiliensis (Müll. Arg.) using the temporary immersion technique. In Vitro Cell Dev Biol Plant 33:81–87CrossRefGoogle Scholar
  14. Hayashi M, Fujiwara K, Kozai T, Tateno M, Kitaya Y (1995) Effects of lighting cycle on daily CO2 exchange and dry weight increase of potato plantlets cultured in vitro photoautotrophically. Acta Hortic 393:213–218Google Scholar
  15. Hilton MG, Wilson PDG (1995) Growth and uptake of sucrose and mineral ions by transformed root cultures of Datura stramonium, Datura candida aurea, Datura wrightii, Hyoscyamus muticus and Atropa belladonna. Planta Medica 61:345–350PubMedCrossRefGoogle Scholar
  16. Kozai T, Iwanami Y, Fujiwara K (1987) Environment control for mass propagation of tissue cultured plantlets. (1) Effects of CO2 enrichment on the plantlet growth during the multiplication stage. Plant Tissue Cult Lett 4:22–26Google Scholar
  17. Kozai T, Fujiwara K, Hyashi M, Aitken-Christie J (1992) The in vitro environment and its control in micropropagation. In: Kurata K, Kozai T (eds) Transplant production systems. Kluwer Academic Publishers, Dordrecht, pp 247–282Google Scholar
  18. Lian ML, Chakrabarty D, Paek KY (2002) Growth and uptake of sucrose and mineral ions by bulblets of Lilium oriental hybrid Casablanca during bioreactor culture. J Hortic Sci Biotechnol 77:253–257 Google Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  20. Paek KY, Chakrabarty D (2003) Micropropagation of woody plants using bioreactor. In: Jain SM, Ishii K (eds) Micropropagation of woody trees and fruits. Kluwer Academic Publishers, Dordrecht, pp 756–766Google Scholar
  21. Paek KY, Chakrabarty D, Hahn EJ (2005) Application of bioreactor system for large scale production of horticultural and medicinal plants. Plant Cell Tissue Organ Cult 81:287–300CrossRefGoogle Scholar
  22. Pareilleux A, Chaubet N (1981) Mass cultivation of Medicago sativa growing on lactose: kinetic aspects. J Appl Microb Biotechnol 11:222–225CrossRefGoogle Scholar
  23. Peavey DG, Steup M, Gibbs M (1977) Characterization of starch breakdown in the intact spinach chloroplast. Plant Physiol 60: 305– 308PubMedCrossRefGoogle Scholar
  24. Pütter J (1974) Peroxidases. In: Bergmeyer HU (eds) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 685–690Google Scholar
  25. Rao AV, Bala K, Tarafdar JC (1990) Dehydrogenase and phosphatase activities in soil as influenced by the growth of arid-land crops. J Agric Sci 115:221–225CrossRefGoogle Scholar
  26. Rufty TW, Israel DW, Volk RJ, Qiu J, Sa T (1993) Phosphate regulation of nitrate assimilation in soybean. J Exp Bot 44:879–891CrossRefGoogle Scholar
  27. Sairam RK, Deshmukh PS, Saxena DC (1998) Role of antioxidant systems in wheat genotypes tolerance to water stress. Biol Plant 41:387–394CrossRefGoogle Scholar
  28. Schenk N, Hsiao KC, Bornman CH (1991) Avoidance of precipitation and carbohydrate breakdown in autoclaved plant tissue culture medium. Plant Cell Rep 10:115–119CrossRefGoogle Scholar
  29. Shin KS, Chakrabarty D, Ko JY, Han SS, Paek KY (2002) Sucrose utilization and mineral nutrient uptake during hairy root growth of red beet (Beta vulgaris L.) in liquid culture. Plant Growth Regul 39:187–193CrossRefGoogle Scholar
  30. Smith IK, Vierheller TL, Thorne CA (1988) Assay of glutathione reductase in crude tissue homogenates using 5,5′-dithiobis(2-nitrobenzoic acid). Anal Biochem 175:408–413 PubMedCrossRefGoogle Scholar
  31. Teisson C, Alvard D (1995) A new concept of plant in vitro cultivation liquid medium: temporary immersion. In: Terzi M (ed) Current issues in plant molecular and cellular biology. Kluwer Academic Publishers, Dordrecht pp 105–110Google Scholar
  32. Tisserat B, Vandercook CE (1986) Computerized-long term tissue culture for orchids. Am Orch Soc Bul 55:35–42 Google Scholar
  33. Zhu LH, Li XY, Welander M (2005) Optimization of growing conditions for the apple rootstock M26 grown in RITA containers using temporary immersion principle. Plant Cell Tissue Organ Cult 81:313–318CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • D. Chakrabarty
    • 1
    • 3
  • Y. H. Dewir
    • 2
    • 3
  • E. J. Hahn
    • 3
  • S. K. Datta
    • 1
  • K. Y. Paek
    • 3
  1. 1.Floriculture SectionNational Botanical Research InstituteLucknowIndia
  2. 2.Department of Horticulture, Faculty of AgricultureKafr El Sheikh UniversityKafr El SheikhEgypt
  3. 3.Research Center for the Development of Advanced Horticultural TechnologyChungbuk National UniversityCheong-juKorea

Personalised recommendations